Novel 2{62 -hydroxy-3{60 -amino-steroids and derivatives thereof and the processes for their preparation

ABSTRACT

The invention relates to novel 2 Beta -hydroxy-3 Alpha -aminosteroids of the formula   , IN WHICH R1 hydrogen or a methyl group R2 an alkyl group having 1-4 carbon atoms R3 hydrogen or an acyl radical X H2, H(OR3) or O, and Y O, H( Beta OR3), R4( Beta OR3), H( Beta COCH3) or H( Beta CHOR3CH3), wherein R4 is a saturated or unsaturated lower alkyl group, their salts and esters and the N-acyl-derivatives thereof, and to the processes for their preparation. The novel compounds are potent anti-arrhythmic agents, devoid of convulsant, local anaesthetic or CNS stimulating activities.

United States Patent Hewett et al.

[ Jan. 21, 1975 NOVEL ZB-HYDROXY-Sa-AMINO-STEROIDS AND DERIVATIVESTHEREOF AND THE PROCESSES FOR THEIR PREPARATION Inventors: Colin LeslieHewett, Bearsden; David Samuel Savage, Newton Mearns, both of ScotlandAkzona Incorporated, Asheville, NC.

Filed: July 10, 1973 Appl. No.: 377,977

Assignee:

Foreign Application Priority Data July 14, 1972 Great Britain 33112/72U.S. Cl 260/397.4, 260/397.45, 260/397.5,

424/243, 260/349 Int. Cl. C07c 169/20, C076 169/32 Field of Search260/397.4, 397.45, 397.5

References Cited UNITED STATES PATENTS 5/1970 Klimstra 260/349 PrimaryExaminerElbert L. Roberts Attorney, Agent, or Firm-Francis W. Young;Philip M. Pippenger; Hugo E. Weisberger [57] ABSTRACT The inventionrelates to novel 2B-hydroxy-3a-aminosteroids of the formula The novelcompounds are potent anti-arrhythmic agents, devoid of convulsant, localanaesthetic or CNS stimulating activities.

3 Claims, No Drawings NOVEL 2B-HYDROXY-3a-AMINO-STEROIDS AND DERIVATIVESTHEREOF AND THE PROCESSES FOR THEIR PREPARATION This invention relatesto novel 2B-hydroxy-3aaminosteroids and salts, esters andN-acyl-derivatives thereof, and the processes for the preparationthereof.

A number of steroids substituted in the 2B-position with a hydroxylgroup and in the 3a-position with a ter tiary amino group are known(British Specification 1,108,563) and some of these have been found topossess anti-arrhythmic activity. However, at therapeutic dose levelsthese compounds also exhibit undesirable activities such as convulsantactivity and local anaesthetic activity which precludes their clinicalapplication. Surprisingly we have discovered that steroids of theandrostane, oestrane and pregnane series substituted in the ZB-positionwith a hydroxyl or acyloxy group and in the 3a-position with a primaryamino group are potent anti-arrhythmic agents which are virtually devoidof convulsant or local anaesthetic activities. Moreover the compounds ofthe invention have no CNS stimulating effects, whereas the correspondingknown tertiary amino compounds have such effects.

These novel primary amino steroids therefore have a great advantage overthe previously described tertiary amino steroids for use asanti-arrhythmic agents.

Therefore, the present invention relates to a novel group of steroids ofthe androstane, oestrane and pregnane series having in 2B-position ahydroxy-, or acyloxy group and in 3a-position a primary amino group, andto salts and N-acyl derivatives thereof. An important and specific groupof compounds according to the present invention are the novel compoundsof the formula:

in which R, hydrogen or a methyl group,

R an alkyl group having 1-4 carbon atoms,

R hydrogen or an acyl radical,

X H H(OR or O, and

Y O, H(BOR R4(BOR3), H(BCOCH or H(BCHOR CH wherein R is a saturated orunsaturated lower alkyl group, and salts and N-acyl-derivatives of thesecompounds.

The compounds according to the invention can be prepared by reacting a2B-hydroxy-3a-haloor 2B- acyloxy-3a-halosteroid of the androstane,oestrane or pregnane series with ammonia, after which the thus obtained2B-hydroxy-3a-aminoor 2B-acyloxy-3aaminocompounds may be converted bymethods known per se into salts thereof, and/or may be N-acylated.

The starting A -compounds are described in the literature, for example,in U.S. Pat. No. 2,940,967; South African patent 64/5098; U.S. Pat. Nos.3,082,223; 3,169,136; 3,338,929; German Pat. No. 1,257,141; Journal Org.Chem. 196i, 3023: and J. Med. Chem. I966, 685.

The reaction is usually carried out by refluxing the mixture for sometime, generally for l to 60 minutes. The solvent may be a neutralsolvent such as dioxan. The starting substances, e.g. the2B-hydroxy-3a-haloor 2B-acyloxy-3a-halo-steroids, can be prepared byreacting a A -steroid of the androstane, oestrane or pregnane serieswith a hypohalous acid, e.g. N-bromo acetamide or N-chloro succinimide,and aqueous perchloric acid followed, if desired, by acylation of the2B- hydroxy-group.

An alternative preparation of the compounds according to the inventionconsists in reacting a ZBJB-oxidosteroid of the androstane, oestrane andpregnane series with ammonia, after which the thus obtained 28-hydroxy-3a-amino compound may be converted by methods known per se into2B-acyloxy-, or ZB-acyloxy- N-acyloxy-derivatives, and/or into salts ofthese compounds.

The amination reaction may be carried out by heating the reactantstogether, usually in the temperature range 60 to 250C; a suitablesolvent such as an alcohol, e.g. ethanol, or an ether, e.g. glycol monoalkyl ether may be used. The reaction may by preference be carried outunder pressure. It may be of advantage to perform the present reactionin the presence of water.

The 2B,3B-oxido-steroids to be used as starting materials may beprepared, for example, from the previously mentionedZB-hydroxy-3a-halo-steroids by treatment with alkali.

A further method for the preparation of the 3aprimary amino steroidsaccording to the present invention consists in reacting a2B,3B-oxido-steroid of the androstane, oestrane and pregnane series withbenzylamine, whereafter the ZB-hydroxyor 2B-acyloxy-3abenzylaminosteroid is submitted to hydrogenolysis e.g. by hydrogenation over asuitable catalyst such as palladium-charcoal.

A further alternative method consists in reacting a 2B,3B-oxido steroidof the androstane, oestrane or pregnane series with an alkali metalazide to give the corresponding 2B-hydroxy-3a-azido steroid which isconverted into the 3B-hydroxy-3a-primary amino steroid by reduction,e.g. with hydrogen in the presence of a metal catalyst. The2B-hydroxy-3a-azido steroid may be acylated first to a2B-acyloxy-3a-azido steroid before reducing to give a2B-acyl0xy-3a-amino-steroid.

The conversion of the Sat-amino compounds into salts thereof can becarried out by treatment with an organic acid such as citric acid,pyruvic acid, succinic acid, fumaric acid and the like, or an inorganicacid such as hydrochloric acid, hydrobromic acid, phosphoric acid.

A 2/3-hydroxy-3a-primary amino steroid may be converted into themono-N-acyl derivative by acylation with an acid anhydride in a suitablesolvent such as methylene chloride; the resulting ZB-hydroxy-N-acyl-3a-amino steroid may be further converted into the 2B-acyloxy-3a-acylamino steroid e.g. by treatment with an acid anhydridewhich may be different from that used to acylate the amino group, inpyridine.

A ZB-acyloxy-Sa-primary amino steroid may be converted into the2B-acyloxy-3a-acylamino-derivatives by treatment with an acid anhydride,which may be different from that present in the 2-position, in asuitable solvent such as methylene chloride.

A l7-oxo-group, if present, is possibly reduced to a l7B-hydroxy groupe.g. with NaBH,,.

Free hydroxyl groups, if present in the molecule, may be acylated.

The acylation may take place with an organic carboxylic acid, preferablywith an aliphatic, cyclo-aliphatic, aromatic or araliphatic carboxylicacid having ll8 carbon atoms, like acetic acid, valeric acid, caproicacid, oenanthic acid, capric acid, lauric acid, benzoic acid, andB-phenyl propionic acid, or with a functional derivative thereof, suchas the anhydrides or the acid chlorides.

A special group of the compounds of the invention is the group ofsteroids having the formula:

in which Z O, H(OH) or COCH R H or acetyl, and the acid-addition saltsand the N-acetyl-derivatives thereof.

The new compounds according to the invention may be applied in the formof pharmaceutical preparation, for which purpose they are mixed withcarriers suitable for oral administration or for injection.

On account of animal experiments the effective oral dose is envisaged asfrom 1-50 mg per kg body weight, and the effective intravenous dose asfrom ().2-l mg per kg body weight.

The following examples illustrate the invention.

EXAMPLE 1 A solution of 70% perchloric acid (60 ml) in water (280 ml)was added to a stirred solution of Sat-androst- 2-en-17-one (200 g) inether (1.15 l) at 15 followed by portionwise addition of N.bromacetamide(112 g) over minutes. After stirring for 1 hour the precipitated whitecrystalline solid was filtered off, washed neutral with ether and waterand crystallised to give 23- hydroxy-3a-bromo-5a-androstan-17-one (172g). The ether layer of the filtrate was washed neutral, dried (Na SO andconcentrated to give a second crop (28 g).

The two crops (200 g) were suspended in methanol (1 l), lON aqueouspotassium hydroxide solution (100 ml) added and the mixture slowlydistilled over 45 minutes. Addition of water precipitated the product asa white solid which was filtered off, washed with water, dried anddissolved inether. Crystallisation from ether: light petrol gave2B,3B-epoxy-5a-androstan-17-one (88 g).

A solution of the 2B,3B-epoxide (30 g) in ethanol (130 ml), water ml)and liquid ammonia was heated in an autoclave at 150 for 6 hours and theresultant crystalline suspension evaporated to dryness. Water (35 ml)and acetic acid (36 ml) were added and the solution kept at 90 for 1hour, cooled and excess water added. The precipitated material wasfiltered off and the filtrate made alkaline with aqueous lON potassiumhydroxide solution to precipitate a white solid which was filtered off,washed neutral with water, dissolved in methylene chloride, the solutiondried (Na S0,), concentrated and ether added to give 25-hydroxy-3a-amino-5a-androstan-l7-one (14.3 g), m.p. l92195C, [01],, +109(in chloroform).

Treatment of 2B-hydroxy-3a-amino-5a-androstan- 17-one with hydrochloricacid in ethanol gave the hydrochloric acid addition salt, m.p. 285C(decomp.); [01],, +104.5C (in ethanol).

The following compounds were prepared in a similar manner:

2B-hydroxy-3a-amino-5aandrostane-l 1,17-dione,28,1la-dihydroxy-3a-amino-5a-androstan-l7-one,2,8,11B-dihydroxy-3a-amino-Sa-androstan-l7-one,2,8-hydroxy-3a'amino-5a-pregnan-20-one, ZB-hydroxy-3a-amino-5a-oestran-1 7-one, 3a-amino-Sa-pregnane-2B,20a-diol,3a-amino-Sa-pregnane-2B,2OB-diol, 3a-amino-5a-oestrane-2B, l 7B-diol,and their hydrochloric acid-addition salts.

EXAMPLE ll A solution of sodium azide (5.5 g) in water (22 ml) was addedto a solution of 2B,3B-epoxy-5a-androstanl7-one (10 g) indimethylacetamide (50 ml) and the boiling solution stirred for 2 hours,cooled and the product crystallised from methylene-dichloride: ether togive 2B-hydroxy-3a-azido-5a-androstan-17-one as a crystalline solid(7.62 g), m.p. 16l165C, [01] +144 (in chloroform).

This azide (5 g) was hydrogenated in methanol (60 ml) over 5% palladiumcharcoal catalyst (0.5 g) and the product crystallised from methylenedichloride: ether to give ZB-hydroxy-3a-amino-5a-androstanl7-one (3.56g) which was identical with an authentic specimen.

EXAMPLE 111 A solution of 2B,3Bepoxy-Sa-androstan-17-one (10 g) in water(10 ml) and benzylamine (40 ml) was boiled under reflux for 5 hours,cooled and the product precipitated as a white solid (13.8 g) by theaddition of water. A solution of this product in acetic acid (25 ml) andwater (25 ml) was heated at for 45 minutes, cooled, water (200 ml) addedand the suspension filtered. The filtrate was made alkaline by thecareful addition of excess aqueous lON potassium hydroxide solution toprecipitate a white solid (13.8 g) which was filtered off, well washedwith water and dried. Crystallisation of this product from methylenedichloride: ether yielded 2B-hydroxy-3a-benzylamino-Sa-androstan- 17-one(10.58 g), m.p. l68173C.

The 3a-benzylamino-2B-ol (4 g) was hydrogenated in ethanol ml) over 5%palladium charcoal catalyst (1.1 g) and the product crystallised frommethylene dichloride: ether to giveZB-hydroxy-Ba-amino-Saandrostan-l7-one (2.2 g) identical with anauthentic specimen.

EXAMPLE IV Sodium borohydride (1.2 g) was added to a solution ofZB-hydroxy-3a-amino-Sa-androstan-17-one (3 g) in methanol (30 ml) andmethylene dichloride (9 ml), the solution stirred for 6 hours and themethylene dichloride evaporated off under reduced pressure at 30.Addition of water precipitated a white solid which was filtered off,washed with water, dried and crystallised from methylene dichloride:ether to give 3a-amino-5aandrostane-2B,l7B-diol in prisms (2.4 g), m.p.

2l8227. Reaction with hydrochloric acid gave the hydrochloricacid-addition salt.

In a similar manner the following compounds were prepared:

3a-amino-5a-androstane-2B,l la,l7B-triol3a-amino-5a-pregnane-2B,20B-diol 3oz-amino-5a-oestrane-2B,l7B-diol.

EXAMPLE V A solution 2B-hydroxy-3a-amino-Sa-androstanl7-one (6 g) inacetic anhydride (25 ml) and pyridine ml) was kept at 90 for 45 minutes,the solution cooled and water added carefully to precipitate a solid.This solid was filtered off, washed neutral with water, dried andcrystallised from methylene dichloride: ether to give2B-acetoxy-3a-acetamido-Sat-androstan-17-one in needles (6.42 g) m.p.227'229.

In a similar manner the following compounds were prepared:

2/3, 1 7B-diacetoxy-3a-acetamido-5a-androstane2B-acetoxy-3wacetamido-Sa-androstane-l l,l7-

dione 23,1la-diacetoxy-3a-acetamido-Sa-androstane- 17-oneZB-acetoxy-Ba-acetamido-Sa-androstan-1 1 8-01- 28,11oz-diacetoxy-3a-acetamido-5a-androstanl7-oneZB-acetoxy-3a-acetamido-Sa-pregnan-ZO-one2B,20B-diacetoxy-3a-acetamido-Sa-pregnaneZB-acetoxy-3a-acetamido-Sa-pregnane1 1,20-dione 213,1la-diacetoxy-Ela-acetamido-Sa-pregnan--one2B-acetoxy-3a-acetamido-Sa-pregnan-l lB-ol- 20-one2B,ZOB-diacetoxy-3a-acetamido-5a-pregnanll-one.

EXAMPLE Vl 2B-Hydroxy-3a-amino-5aandrostan-17-one (2.5 g) was added toacetic anhydride (10 ml) at 20 and the reaction stirred for 10 minutes;almost immediately a product began to crystallise out. Water was addedto precipitate completely a white solid (2.43 g) which was filtered off,washed with water, dried and crystallised from methylene dichloride:ether to give ZB-hydroxy- 3a-acetamido-5a-androstan-l7-one in prisms(2.28 g), m.p. 239-243.

In a similar manner the following compounds were prepared:

2B-hydroxy-3a-acetamido-Sa-androstane-l l, l 7- dione 213,11a-dihydroxy-3a-acetamido-Sa-androstanl7-one 2B,]lB-dihydroxy-3a-acetamido-Sa-androstanl7-oneZB-hydroxy-3a-acetamido-Sa-pregnan-ZO-one2B-hydroxy-3u-acetamido-Sa-pregnane-l l,20-dione 2B,]la-dihydroxy-3a-acetamido-Sa-pregnan-ZO-one 28,1lB-dihydroxy-3a-acetamido-Sa-pregnan- 20-one.

EXAMPLE Vll A solution of 2B-hydroxy-3a-azidoSa-androstanl7-one (5 g) inacetic anhydride (10 ml) and pyridine (10 ml) was heated at for 45minutes. The product was worked up in the usual manner and crystallisedfrom hexane to give 2B-acetoxy-3a-azido-Saandrostan-l7-one (4.4 g), m.p.l07-l09.

This azide (2 g) was hydrogenated in ethanol (20 ml) over 5% palladium:charcoal catalyst (0.5 g) to give 23-acetoxy-3a-amino-5a-androstan-l7-0ne which crystallised from ether.

in a similar manner the following compounds and their hydrochloridesalts were prepared:

2B-acetoxy-3a-amino-5 a-androstane'l l,l7-dione 2,8,11a-diacetoxy-3a-amino-5a-androstan-17-one2B-acetoxy-3a-amino-5a-androstan-l lB-ol-l 7-one2B-acetoxy-3a-amino-5a-pregnan-ZO-one 2B-acetoxy-3a-amino-5 a-oestranl7-one.

What is claimed is:

l. A 2B-hydroxy-3a-amino-steroid of the group consisting of theoestrane, androstane and pregnane series,

of the formula:

R Y A l RI a H Nn H H in which R, is selected from the group consistingof hydrogen and methyl;

R is methyl;

R is selected from the group consisting of hydrogen and acyl derivedfrom an aliphatic, cycloaliphatic, aromatic or araliphatic carboxylichaving l-l 8 carbon atoms;

X is selected from the group consisting of H H(OR and O;

Y is selected from the group consisting of 0,

H(BOR H(BCOCH and H(flCHOR CH and the pharmaceutically acceptable acidaddition salts and N-mono-acyl derivatives thereof.

2. A 2-B-hydroxy-3wamino-steroid of the formula:

in which Z is selected from the group consisting of O, H(OH) and COCH Ris selected from the group consisting of H and acetyl; and thepharmaceutically acceptable acid addition salts and N-acetyl derivativesthereof.

3. A member of the group consisting of ZB-hydroxy-3a-amino-5a-androstan-l7-one and its hydrochloric acid addition salt.

2. A 2- Beta -hydroxy-3 Alpha -amino-steroid of the formula:
 3. A memberof the group consisting of 2 Beta -hydroxy-3 Alpha -amino-5 Alpha-androstan-17-one and its hydrochloric acid addition salt.